Oleosin Is Bifunctional Enzyme That Has Both Monoacylglycerol Acyltransferase and Phospholipase Activities

In plants, fatty oils are generally stored in spherical intracellular organelles referred to as oleosomes that are covered by proteins such as oleosin. Seeds with high oil content have more oleosin than those with low oil content. However, the exact role of oleosin in oil accumulation is thus far unclear. Here, we report the isolation of a catalytically active 14 S multiprotein complex capable of acylating monoacylglycerol from the microsomal membranes of developing peanut cotyledons. Microsomal membranes from immature peanut seeds were solubilized using 8 M urea and 10 mM CHAPS. Using two-dimensional gel electrophoresis and mass spectrometry, we identified 27 proteins in the 14 S complex. The major proteins present in the 14 S complex are conarachin, the major allergen Ara h 1, and other seed storage proteins. We identified oleosin 3 as a part of the 14 S complex, which is capable of acylating monoacylglycerol. The recombinant OLE3 microsomes from Saccharomyces cerevisiae have been shown to have both a monoacylglycerol acyltransferase and a phospholipase A(2) activity. Overexpression of the oleosin 3 (OLE3) gene in S. cerevisiae resulted in an increased accumulation of diacylglycerols and triacylglycerols and decreased phospholipids. These findings provide a direct role for a structural protein (OLE3) in the biosynthesis and mobilization of plant oils.

Serine/Threonine/Tyrosine Protein Kinase Phosphorylates Oleosin, a Regulator of Lipid Metabolic Functions

Plant oils are stored in oleosomes or oil bodies, which are surrounded by a monolayer of phospholipids embedded with oleosin proteins that stabilize the structure. Recently, a structural protein, Oleosin3 (OLE3), was shown to exhibit both monoacylglycerol acyltransferase and phospholipase A(2) activities. The regulation of these distinct dual activities in a single protein is unclear. Here, we report that a serine/threonine/tyrosine protein kinase phosphorylates oleosin. Using bimolecular fluorescence complementation analysis, we demonstrate that this kinase interacts with OLE3 and that the fluorescence was associated with chloroplasts. Oleosin-green fluorescent protein fusion protein was exclusively associated with the chloroplasts. Phosphorylated OLE3 exhibited reduced monoacylglycerol acyltransferase and increased phospholipase A(2) activities. Moreover, phosphatidylcholine and diacylglycerol activated oleosin phosphorylation, whereas lysophosphatidylcholine, oleic acid, and Ca2+ inhibited phosphorylation. In addition, recombinant peanut (Arachis hypogaea) kinase was determined to predominantly phosphorylate serine residues, specifically serine-18 in OLE3. Phosphorylation levels of OLE3 during seed germination were determined to be higher than in developing peanut seeds. These findings provide direct evidence for the in vivo substrate selectivity of the dual-specificity kinase and demonstrate that the bifunctional activities of oleosin are regulated by phosphorylation.

Development of micro-shock wave assisted dry particle and fluid jet delivery system

Small quantity of energetic material coated on the inner wall of a polymer tube is proposed as a new method to generate micro-shock waves in the laboratory. These micro-shock waves have been harnessed to develop a novel method of delivering dry particle and liquid jet into the target. We have generated micro-shock waves with the help of reactive explosive compound high melting explosive (octahydro-1,3,5,7-tetranitro-1,3,5,7-tetrazocine) and traces of aluminium] coated polymer tube, utilising 9 J of energy. The detonation process is initiated electrically from one end of the tube, while the micro-shock wave followed by the products of detonation escape from the open end of the polymer tube. The energy available at the open end of the polymer tube is used to accelerate tungsten micro-particles coated on the other side of the diaphragm or force a liquid jet out of a small cavity filled with the liquid. The micro-particles deposited on a thin metal diaphragm (typically 100-mu m thick) were accelerated to high velocity using micro-shock waves to penetrate the target. Tungsten particles of 0.7 mu m diameter have been successfully delivered into agarose gel targets of various strengths (0.6-1.0 %). The device has been tested by delivering micro-particles into potato tuber and Arachis hypogaea Linnaeus (ground nut) stem tissue. Along similar lines, liquid jets of diameter 200-250 mu m (methylene blue, water and oils) have been successfully delivered into agarose gel targets of various strengths. Successful vaccination against murine salmonellosis was demonstrated as a biological application of this device. The penetration depths achieved in the experimental targets are very encouraging to develop a future device for biological and biomedical applications.

Grafting of Dispersants on MoS2 Nanoparticles in Base Oil Lubrication of Steel

Solid lubricant nanoparticles in suspension in oil are good lubricating options for practical machinery. In this article, we select a range of dispersants, based on their polar moieties, to suspend 50-nm molybdenum disulfide particles in an industrial base oil. The suspension is used to lubricate a steel on steel sliding contact. A nitrogen-based polymeric dispersant (aminopropyl trimethoxy silane) with a free amine group and an oxygen-based polymeric dispersant (sorbital monooleate) when grafted on the particle charge the particle negatively and yield an agglomerate size which is almost the same as that of the original particle. Lubrication of the contact by these suspensions gives a coefficient of friction in the similar to 0.03 range. The grafting of these surfactants on the particle is shown here to be of a chemical nature and strong as the grafts survive mechanical shear stress in tribology. Such grafts are superior to those of other silane-based test surfactants which have weak functional groups. In the latter case, the particles bereft of strong grafts agglomerate easily in the lubricant and give a coefficient of friction in the 0.08-0.12 range. This article investigates the mechanism of frictional energy dissipation as influenced by the chemistry of the surfactant molecule.

AN EXPERIMENTAL STUDY ON EFFERVESCENT ATOMIZATION OF BIO-OIL FUELS

This paper presents the work on detailed characterization of effervescent spray of Jatropha and Pongamia pure plant oils. The spray characteristics of these biofuels are compared with those of diesel. Both macroscopic and microscopic spray characteristics at different injection pressures and gas-to-liquid ratio (GLR) have been studied. The particle/droplet imaging analysis (PDIA) technique along with direct imaging methods are used for the purpose of spray characterization. Due to their higher viscosity, pure plant oils showed poor atomization compared to diesel and a blend of diesel and pure plant oil at a given GLR. Pure plant oil sprays showed a lower spray cone angle when compared to diesel and blends at lower GLRs. However, the difference is not significant at higher GLRs. Droplet size measurements at 100 mm downstream of the exit orifice showed reduction in Sauter mean diameter (SMD) diameter with increase in GLR. A radial variation in the SMD is observed for the blend and pure plant oils. Pure oils showed a larger variation when compared to the blend. Spray unsteadiness has been characterized based on the image-to-image variation in the mean droplet diameter and fluctuations in the spray cone angle. Results showed that pure plant oil sprays are more unsteady at lower GLRs when compared to diesel and blend. A critical GLR is identified at which the spray becomes steady. The three regimes of spray operation, namely ``steady spray,'' ``pulsating spray,'' and ``spray and unbroken liquid jet'' are identified in the injection pressure-GLR parameter space for these pure plant oils. Two-phase flow imaging inside the exit orifice shows that for the pure plant oils, the flow is highly transient at low GLRs and the bubbly, slug, and annular two-phase flow regimes are all observed. However, at higher GLRs where the spray is steady, only the annular flow regime is observed.

FLOTATION OF QUARTZ WITH SOAPS OF BOMBAX MALABARICA OIL AND SHARK LIVER OIL AS COLLECTORS

An attempt has been made to use the indigeneous oils-Bombax Malabarica oil and Shark liver oil in the form of sodium soaps as collectors in the flotation of quartz using barium chloride as activator. The effect of pH, collector concentration and activator concentration on the flotation of quartz is studied in a Leaf and knoll flotation cell. The experiments show that it is possible to obtain 98.0 per cent of quartz as float using 10 mg. of Bombax. Malabarica oil and Shark liver oil soaps at pH values of more than 7.0 and when barium ion concentration is in excess of that required to form barium soaps. Bombax Malabarica oil is found to be superior to Shark liver oil as collector in the flotation of quartz.

SWEETENING OF BIOGAS IN SPRAY TOWERS FOR POWER GENERATION

This paper presents the experience of the new design of using impinging jet spray columns for scrubbing hydrogen sulfide from biogas that has been developed by Indian Institute of Science and patented. The process uses a chelated polyvalent metal ion which oxidizes the hydrogen sulfide to sulfur as a precipitate. The sulfur generated is filtered and the scrubbing liquid recycled after oxidation. The process involves in bringing contact the sour gas with chelated liquid in the spray columns where H2S reacts with chelated Fe3+ and precipitates as sulfur, whereas Fe3+ gets reduced to Fe2+. Fe2+ is regenerated to Fe3+ by reaction of oxygen in air in a separate packed column. The regenerated liquid is recirculated. Sulfur is filtered and separated as a byproduct. The paper presents the experience in using the spray towers for hydrogen sulfide removal and further use of the clean gas for generating power using gas engines. The maximum allowable limit of H2S for the gas engine is 200 ppm (v/v) in order to prevent any corrosion of engine parts and fouling of the lubricating oil. With the current ISET process, the hydrogen sulfide from the biogas is cleaned to less than 100 ppm (v/v) and the sweet gas is used for power generation. The system is designed for 550 NM3/hr of biogas and inlet H2S concentration of 2.5 %. The inlet concentration of the H2S is about 1 - 1.5 % and average measured outlet concentration is about 30 ppm, with an average gas flow of about 300 - 350 NM3/hr, which is the current gas production rate. The sweet gas is used for power generation in a 1.2 MWe V 12 engine. The average power generation is about 650 - 750 kWe, which is the captive load of the industry. The plant is a CHP (combined heat power) unit with heat from the cylinder cooling and flue being recovered for hot water and steam generation respectively. The specific fuel consumption is 2.29 kWh/m(3) of gas. The system has been in operation for more than 13,000 hours in last one year in the industry. About 8.4 million units of electricity has been generated scrubbing about 2.1 million m3 of gas. Performance of the scrubber and the engine is discussed at daily performance level and also the overall performance with an environment sustenance by precipitating over 27 tons of sulfur.

EXPERIMENTAL STUDIES ON AIR-ASSISTED ATOMIZATION OF JATROPHA PURE PLANT OIL

The present study focuses on exploring air-assisted atomization strategies for effective atomization of high-viscosity biofuels, such as pure plant oils (PPOs). The first part of the study concerns application of a novel air-assisted impinging jet atomization for continuous spray applications, and the second part concerns transient spray applications. The particle/droplet imaging analysis (PDIA) technique along with direct imaging methods are used for the purpose of spray characterization. In the first part, effective atomization of Jatropha PPO is demonstrated at gas-to-liquid ratios (GLRs) on the order 0.1. The effect of liquid and gas flow rates on the spray characteristics is evaluated, and results indicate a Sauter mean diameter (SMD) of 50 mu m is achieved with GLRs as low as 0.05. In the second part of the study, a commercially available air-assisted transient atomizer is evaluated using Jatropha PPO. The effect of the pressure difference across the air injector and ambient gas pressure on liquid spray characteristics is studied. The results indicate that it is possible to achieve the same level of atomization of Jatropha as diesel fuel by operating the atomizer at a higher pressure difference. Specifically, a SMD of 44 mu m is obtained for the Jatropha oil using injection pressures of <1 MPa. A further interesting observation associated with this injector is the near constancy of a nondimensional spray penetration rate for the Jatropha oil spray.